By Myrtle A. Davis
Dr. Myrtle A. Davis has assembled a panel of state of the art scientists to explain their top equipment for detecting, illuminating, and quantifying apoptotic mechanisms in a fashion that's important for the layout of toxicology and pharmacology reviews. those cutting-edge recommendations comprise move cytometric, fluorometric, and laser scanning tools for quantifying and characterizing apoptosis, in addition to protocols for using DNA microarray know-how, excessive throughput displays, and ELISA. Immunocytochemical equipment for measuring biochemical and molecular endpoints in tissue sections should be hugely precious for these accomplishing reports in entire animal versions instead of telephone tradition structures.
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Dr. Myrtle A. Davis has assembled a panel of state of the art scientists to explain their top equipment for detecting, illuminating, and quantifying apoptotic mechanisms in a manner that's invaluable for the layout of toxicology and pharmacology reports. those cutting-edge suggestions contain circulation cytometric, fluorometric, and laser scanning equipment for quantifying and characterizing apoptosis, in addition to protocols for using DNA microarray expertise, excessive throughput monitors, and ELISA.
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Additional resources for Apoptosis Methods in Pharmacology and Toxicology: Approaches to Measurement and Quantification (Methods in Pharmacology and Toxicology)
2000) Apoptosis, bcl-2 expression, and proliferation in benign and malignant endometrial epithelium: An approach using multiparameter flow cytometry. Gynecol Oncol. 77(1), 11–17. Apoptotic Cell Death 35 66. , Ramaekers, F. , and Reutelingsperger, C. P. (1996) A novel assay to measure loss of plasma membrane asymmetry during apoptosis of adherent cells in culture. Cytometry 24(2), 131–139. 67. , Melamed, M. , and Darzynkiewicz, Z. (1999) Analysis of apoptosis by laser scanning cytometry. Cytometry 35(3), 181–195.
Combined use of flow cytometry/cell sorting and confocal laser scanning microscopy. TNF-_/ActD treated Hepa-1 cells were stained with CMX Rosamine and then analyzed for mitochondrial membrane potential and NAD(P)H fluorescence (A). Cells in different regions of the cytogram were then sorted, and subsequently stained with the DNA fluorochrome Hoechst 33342. These cells were then examined by CLSM. (B), (C), and (D) show three-dimensional reconstructions of nuclei from cells sorted from healthy, early apoptotic, and late apoptotic populations.
Totowa, NJ 37 38 Darzynkiewicz et al. nates of cell location on the slide are recorded in a list-mode fashion together with other measured cell parameters, cells can be relocated after the measurement. They can be then examined visually or subjected to image analysis to correlate the observed change in the measured parameter with the change in their morphology. The possibility of cell relocation and other attributes of LSC which are presented further in this chapter, has contributed to this instrument’s usefulness in numerous applications in analysis of apoptosis (10).